title: Genetically-engineered mesenchymal stem cells transfected with human HCN1 gene to create cardiac pacemaker cells.
abstract: OBJECTIVE
  To test the proof-of-principle that genetically-engineered mesenchymal stem cells (MSCs) transfected with the human hyperpolarization-activated cyclic nucleotide-gated channel 1 (hHCN1) gene can be modified to become cardiac pacemaker cells.
  
  
  METHODS
  MSCs were transfected with the hHCN1 gene using lentiviral-based transfection. The expressed pacemaker current (I(f)) in hHCN1-transfected MSCs was recorded using whole-cell patch-clamp analysis. The effect of the hHCN1-transfected MSCs on cardiomyocyte excitability was determined by coculturing the MSCs with neonatal rabbit ventricular myocytes (NRVM). The spontaneous action potentials of the NRVM were recorded by whole-cell current-clamp analysis.
  
  
  RESULTS
  A high level time- and voltage-dependent inward hyperpolarization current that was inhibited by 4 mM caesium chloride was detected in hHCN1-transfected MSCs, suggesting that the HCN1 proteins acted as I(f) channels in MSCs. The mean ± SE beating frequency in NRVMs cocultured with control MSCs transfected with the pcDNA3 plasmid control was 82 ± 8 beats/min (n = 5) compared with 129 ± 11 beats/min (n = 5) in NRVMs cocultured with hHCN1-transfected MSCs.
  
  
  CONCLUSIONS
  Genetically-engineered MSCs transfected with the hHCN1 gene can be modified to become cardiac pacemaker cells.
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China.
authors.forename: Ya-Feng
authors.initials: YF
authors.lastname: Zhou
authors.forename: Xiang-Jun
authors.initials: XJ
authors.lastname: Yang
authors.forename: Hong-Xia
authors.initials: HX
authors.lastname: Li
authors.forename: Lian-Huan
authors.initials: LH
authors.lastname: Han
authors.forename: Wen-Ping
authors.initials: WP
authors.lastname: Jiang
mesh_terms: D000200:Action Potentials; D000818:Animals; D000831:Animals, Newborn; D060846:Cell Engineering / Q000379:methods*; D002586:Cesium / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology; D018920:Coculture Techniques; D015870:Gene Expression; D005822:Genetic Vectors; D006352:Heart Ventricles / Q000166:cytology / Q000187:drug effects; D006801:Humans; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels / Q000235:genetics* / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010957:Plasmids; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D014162:Transfection
chemical_list: D002712:Chlorides; C578665:HCN1 protein, human; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D015221:Potassium Channels; D002586:Cesium; C028019:cesium chloride
keywords:   hHCN1 gene; Cardiac arrhythmias; cardiac pacemaker cells; genetic engineering; mesenchymal stem cells